Abstract
BackgroundThe dual Na+ and cardiac Ca2+-release channel inhibitor, Flecainide (FLEC) is effective in patients with catecholaminergic polymorphic ventricular tachycardia (CPVT), a disease caused by mutations in cardiac Ca2+-release channels (RyR2), calsequestrin (Casq2), or calmodulin. FLEC suppresses spontaneous Ca2+ waves in Casq2-knockout (Casq2−/−) cardiomyocytes, a CPVT model. However, a report failed to find FLEC efficacy against Ca2+ waves in another CPVT model, RyR2-R4496C heterozygous mice (RyR2R4496C+/−), raising the possibility that FLEC efficacy may be mutation dependent.ObjectiveTo address this controversy, we compared FLEC in Casq2−/− and RyR2R4496C+/− cardiomyocytes and mice under identical conditions.MethodsAfter 30 min exposure to FLEC (6 μM) or vehicle (VEH), spontaneous Ca2+ waves were quantified during a 40 s pause after 1 Hz pacing train in the presence of isoproterenol (ISO, 1 μM). FLEC efficacy was also tested in vivo using a low dose (LOW: 3 mg/kg ISO + 60 mg/kg caffeine) or a high dose catecholamine challenge (HIGH: 3 mg/kg ISO + 120 mg/kg caffeine).ResultsIn cardiomyocytes, FLEC efficacy was dependent on extracellular [Ca2+]. At 2 mM [Ca2+], only Casq2−/− myocytes exhibited Ca2+ waves, which were strongly suppressed by FLEC. At 3 mM [Ca2+] both groups exhibited Ca2+ waves that were suppressed by FLEC. At 4 mM [Ca2+], FLEC no longer suppressed Ca2+ waves in both groups. Analogous to the results in myocytes, RyR2R4496C+/− mice (n = 12) had significantly lower arrhythmia scores than Casq2−/− mice (n = 9), but the pattern of FLEC efficacy was similar in both groups (i.e., reduced FLEC efficacy after HIGH dose catecholamine challenge).ConclusionFLEC inhibits Ca2+ waves in RyR2R4496C+/− cardiomyocytes, indicating that RyR2 channel block by FLEC is not mutation-specific. However, FLEC efficacy is reduced by Ca2+ overload in vitro or by high dose catecholamine challenge in vivo, which could explain conflicting literature reports.
Highlights
MATERIALS AND METHODSCatecholaminergic polymorphic ventricular tachycardia (CPVT)is an inherited arrhythmogenic syndrome characterized by polymorphic ventricular tachycardia in response to exercise or emotional stress, leading sometimes to sudden cardiac death (Huikuri et al, 2001)
In approximately 60% of cases, catecholaminergic polymorphic ventricular tachycardia (CPVT) is caused by mutations in three genes encoding sarcoplasmic reticulum (SR) Ca2+ handling proteins, the cardiac ryanodine receptor (RyR2), calsequestrin 2 (Casq2) (Faggioni et al, 2012), or calmodulin (CaM) (Nyegaard et al, 2012; Crotti et al, 2013)
To ensure optimal experimental conditions, we first established the time-dependence of Ca2+ wave suppression by flecainide in single Casq2−/− cardiomyocytes (Figure 1)
Summary
The dual Na+ and cardiac Ca2+-release channel inhibitor, Flecainide (FLEC) is effective in patients with catecholaminergic polymorphic ventricular tachycardia (CPVT), a disease caused by mutations in cardiac Ca2+-release channels (RyR2), calsequestrin (Casq2), or calmodulin. FLEC suppresses spontaneous Ca2+ waves in Casq2-knockout (Casq2−/−) cardiomyocytes, a CPVT model. A report failed to find FLEC efficacy against Ca2+ waves in another CPVT model, RyR2-R4496C heterozygous mice (RyR2R4496C+/−), raising the possibility that FLEC efficacy may be mutation dependent
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